Hg~(2+)胁迫对浮萍体细胞DNA一级结构和抗氧化酶系的损伤(英文)

主要从DNA一级结构及抗氧化酶系变化两方面 ,研究了Hg2 + 胁迫下浮萍体细胞的损伤。结果表明 :运用随机扩增多态性DNA法 (RandomamplifiedpolymorphismDNA ,RAPD)和DNA梯法 (DNALadder) ,5～ 1 0mg·L- 1 Hg2 +处理组可检测到基因组DNA的明显损伤 ,2 0mg·L- 1 Hg2 + 已导致细胞坏死 ;RAPD法较DNALadder法更灵敏。本文还发现 ,活性氧和抗氧化酶系很可能参与了浮萍体细胞凋亡过程。低浓度的Hg2 + 胁迫可刺激抗氧化酶活性升高 ,以清除体内活性氧 ,而一旦活性氧水平超出一定域值 ,抗氧化酶活性急速下降 ,导致细胞凋亡     

Cd2+胁迫下浮萍叶细胞核超微结构、nDNA损伤及抗氧化酶系的变化

以不同浓度Cd2+处理6d的浮萍为材料,从细胞核超微结构损伤、nDNA一级结构变化及抗氧化酶系活性改变等方面分析重金属的植物毒理学效应。透射电镜观察发现,5～7mg/L Cd2+处理的细胞核膜完整,染色质凝聚并趋边化;10mg/L Cd2+处理后则核仁解体,且DNA原位末端标记(TUNEL)检测发现DNA的3′-OH断端可被特异标记,表现出典型的凋亡细胞特征。而20mg/L Cd2+已导致细胞坏死。同时,较低浓度的Cd2+胁迫可刺激抗氧化酶(SOD,POD,CAT)活性升高,以清除体内活性氧,而随金属浓度的进一步增高,三种抗氧化酶活性都急速下降。研究发现,活性氧和抗氧化酶系密切参与了重金属Cd2+诱导的浮萍体细胞凋亡过程。     

Cu2+对拟南芥根的局部毒性及诱导DNA损伤和细胞死亡

该文探讨了不同浓度的Cu²⁺胁迫对拟南芥(Arabidopsis thaliana)根生长、活性氧(ROS)积累、抗氧化酶活性、质膜完整性和细胞活性的影响, 通过分根实验初步分析了Cu²⁺毒性效应的影响范围。结果表明, Cu²⁺胁迫可显著抑制拟南芥主根伸长, 诱导ROS积累及DNA损伤, 促发抗氧化酶活性升高, 破坏质膜完整性, 且Cu²⁺浓度越高, 毒性效应越明显, 在高浓度Cu²⁺胁迫下细胞活性显著降低。分析各参数之间的关系, 表明ROS的积累与超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)及抗坏血酸过氧化物酶(APX)的活性呈显著正相关; ROS积累与DNA损伤、质膜完整性、细胞活性之间具有显著的近线性关系。分根实验结果表明, 只有在添加重金属Cu²⁺(75 μmol·L^–1)一侧培养基中的根生长受抑制, 并出现ROS积累、细胞死亡, 暗示Cu²⁺对拟南芥根系的局部毒性效应可能是由于ROS的局部性积累导致受胁迫根系一侧的细胞死亡所引起的。     

水分胁迫引起的两种不同生态型芦苇的DNA损伤与修复

利用DNA解链荧光分析（FADU）法检测两种不同生态型芦苇（Phragmites communis T.)在PEG6000胁迫处理后的DNA损伤。结果表明：无论是20％还是30％PEG6000胁迫处理,耐旱性强的沙丘芦苇的DNA损伤都比耐旱性弱的沼泽芦苇较轻。利用不同浓度的二乙基二硫代谢氨基甲酸钠（DDC）、H2O2、FeSO4以增加芦苇的3种活性氧（O2^-、H2O2、OH）的实验也同样显示出沙丘芦苇抵抗水分胁迫引起的DNA损伤的能力较强。同时,当加入外源活性氧清除剂二甲基亚砜（DMSO）、抗坏血酸（Vc）时,水分胁迫处理的芦苇DNA损伤表现出不同程度的减轻。当PEG胁迫处理的芦苇复水后,DNA损伤随复水时间延长而逐渐减轻,但沙丘芦苇的DNA损伤修复较快而完全。实验初步证明：水分胁迫可引起植物体内DNA损伤且该损伤与活性氧有关,植物的抗旱性与DNA损伤及修复密切相关。     

钙抑制剂对机械损伤胁迫下合作杨叶片活性氧代谢的影响

以1年生合作杨扦插苗为材料,研究了叶面喷施Ca2+通道阻断剂氯化镧(LaCl3)和Ca2+螯合剂EGTA预处理对机械损伤胁迫下合作杨叶片抗氧化酶活性、过氧化氢(H2O2)和丙二醛(MDA)含量以及氧自由基(O2?-)产生速率的影响.结果显示,与对照相比,机械损伤胁迫下合作杨叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)等抗氧化酶活性均显著升高,活性氧水平和MDA含量显著增加;外源喷施EGTA和LaCl3降低了机械损伤胁迫下叶片SOD、POD、CAT和APX活性,减缓了O2?-产生速率,H2O2含量和MDA含量显著下降;且EGTA的抑制作用比LaCl3更强.研究表明,机械损伤胁迫诱导的活性氧代谢需要Ca2+的参与,Ca2+和活性氧在植物防御信号传递过程中密切相关;伤害诱导胞外Ca2+内流是胞内Ca2+浓度增加的重要来源.     

汞胁迫对白骨壤(Avicennia marina)幼苗生理生态的影响

为了解红树植物的重金属抗性机制,对白骨壤(Avicennia marina)幼苗进行不同浓度Hg2+(1、5、10、50、100 mg·L-1)的胁迫实验,测定并分析了Hg2+胁迫对白骨壤幼苗叶片的光合作用和抗氧化酶活性的影响.结果表明:叶片净光合速率随着胁迫时间的延长而降低,高浓度(≥150 mg·L-1)Hg2+胁迫下叶片的净光合速率低于中低浓度胁迫,且高浓度胁迫的叶片净光合速率在48 h后快速下降;叶片净光合速率与胞间二氧化碳浓度呈极显著负相关,叶绿素含量随Hg2+浓度的增加而降低.气孔导度在不同浓度胁迫下反应不同,低浓度Hg2+对白骨壤幼苗光合的影响可能是气孔因素,中高浓度Hg2+对白骨壤幼苗光合作用的抑制主要是非气孔因素.低浓度Hg2+胁迫,白骨壤幼苗叶片SOD、POD活性升高,表现了一定的抗逆性,而高浓度表现为抑制作用,基本在100 mg·L-1 Hg2+胁迫下活性达到最低值.说明Hg2+可以抑制白骨壤叶片的光合活性,高浓度Hg2+胁迫削弱了白骨壤的活性氧清除能力,植物极易受到伤害.     

外源亚精胺对盐胁迫下马铃薯幼苗生长和生理生化特征的影响

盐胁迫是影响作物生长的主要非生物胁迫类型，引起离子毒害和渗透胁迫，导致植物生长减弱、失绿、萎蔫甚至死亡。前期研究表明，适宜浓度的外源亚精胺能够缓解盐胁迫条件下植物叶片受损伤程度，提升生物膜抵抗盐离子伤害的能力，促进植物生长。该试验采用营养液培养法，以100mmol·L-1、200mmol·L-1、300 mmol· L-1NaCl溶液模拟不同盐胁迫程度，以中度耐盐品种晋薯16号、轻度耐盐品种冀张薯12号为试材，当马铃薯脱毒幼苗长至 4～5 片真叶时，连续叶面喷施0.9mmol·L-1外援亚精胺 7 d，2次/d。分析叶面喷施外源亚精胺（Spd）对不同盐胁迫程度条件下马铃薯幼苗生长、叶片抗氧化酶活性、渗透调节物质含量的影响。结果表明：（1）叶面喷施Spd缓解了盐胁迫对幼苗生长的抑制作用，提高了叶绿素含量和根系活力，提升超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）、抗坏血酸过氧化物酶（APX）、抗坏血酸(ASA)和谷胱甘肽(GSH)等抗氧化酶活性，以及脯氨酸、可溶性糖、氨基酸含量；（2）200 mmol· L-1NaCl胁迫条件下，Spd对“晋薯16号”缓解作用最显著。研究表明Spd通过提高马铃薯幼苗根系活力、叶绿素含量、抗氧化酶活性、渗透调节能力，提高马铃薯幼苗对盐胁迫的适应性，促进马铃薯幼苗生长。     

Cu胁迫对狭叶香蒲体内元素吸收分配的影响

采用水培法,研究了不同浓度Cu²⁺胁迫下狭叶香蒲（Typha angustifolia）体内Cu及其他元素吸收分配的变化。结果表明：狭叶香蒲地上部和根系累积的Cu量均随Cu²⁺浓度升高显著增加;在同一浓度Cu²⁺胁迫下,根中Cu的累积量明显高于地上部。根系同一组织中的Cu百分含量随Cu²⁺浓度的升高均显著增加;同一Cu²⁺浓度下,根系各组织中Cu含量均表现为：中柱>皮层>表皮。低浓度（1和5 mg·L^-1）Cu²⁺对地上部及根中Ca、Mg含量及地上部Mn含量均无显著影响,但高浓度（80和100 mg·L^-1）Cu²⁺可促进植株对Ca的吸收,显著降低对Mg﹑Mn和Zn的吸收。较高浓度（55～100 mg·L^-1）Cu²⁺胁迫后,根系各组织对Ca的吸收增加,对Zn的吸收降低;55～80 mg·L^-1 Cu²⁺胁迫下根系不同组织对Cl和K的吸收增加,但高浓度（100 mg·L^-1）Cu²⁺胁迫下则显著降低。根系累积的Ca、Zn和K主要分配到中柱中,Cl主要分配到皮层中.     

Hg2+对南极冰藻Chlorophyceae L-4生长和抗氧化系统的影响及藻体富集Hg2+的规律

南极冰藻Chlorophyceae L-4是南极生态系统重要的初级生产力和组成部分,其长期生长在极地环境中,有着特殊的生理机制。在生存环境和生长条件发生变化时,冰藻的膜脂系统和蛋白含量都会发生变化,而在受到重金属胁迫时,冰藻的超微结构也会发生明显变化。【目的】研究Chlorophyceae L-4在重金属离子Hg2+胁迫条件下的状态和Hg2+富集以及对其抗氧化系统的影响,为南极环境监测提供依据。【方法】绘制南极冰藻细胞在重金属离子Hg2+不同浓度胁迫条件下的生长曲线,观察其超微结构;测定丙二醛含量和SOD酶活性变化;ICP-MS法研究藻体富集Hg2+规律。【结果】Hg2+在低浓度时(≤100μg/L),细胞个数较正常条件明显偏少;在高浓度时(≥250μg/L),出现细胞死亡。丙二醛含量随Hg2+浓度升高而升高,SOD酶活性则先增强再减弱。藻体富集Hg2+在1 h达到峰值,而在Hg2+浓度持续升高时,富集量轻微降低。【结论】Hg2+离子对冰藻生长有抑制毒害作用;对Chlorophyceae L-4抗氧化系统有明显不利影响;L-4富集Hg2+在1 h内饱和,Hg2+过高时,富集量稍微降低。     

模拟酸雨对乐东拟单性木兰幼苗部分生理指标的影响

采用盆栽法,研究了不同pH值的模拟酸雨胁迫对乐东拟单性木兰（Paramecia lotungensis）叶片膜损伤、抗氧化酶活性、叶绿素（Chl）含量以及气体交换参数等的变化。结果表明：在模拟酸雨胁迫下,幼苗叶片膜透性和丙二醛（MDA）含量显著增加,且两者呈现显著正相关;过氧化氢酶（CAT）活性逐渐下降,超氧化物歧化酶（SOD）和过氧化物（POD）活性逐渐或急剧上升;Chl含量明显下降;净光合速率（Pn）、气孔导度（Ga）、蒸腾速率（Tt）、水分利用效率（WUE）和气孔限制值（La）等指标均呈下降趋势,而胞间CO2浓度（Ci）却表现为上升趋势。表明酸雨诱导了活性氧生成,使抗氧化酶系统失调,造成活性氧产生和清除之间的不平衡,从而加速膜脂过氧化作用,破坏膜透性,降解叶绿素,导致光合作用下降。     

Cytoprotective activity of annphenone against oxidative stress-induced apoptosis in V79-4 lung fibroblast cells

We have elucidated the cytoprotective effect of annphenone (2,4-dihyroxy-6-methoxy-acetophenone 4-O-beta-d-glucopyranoside) against oxidative stress-induced apoptosis. Annphenone scavenged intracellular reactive oxygen species (ROS) and increased antioxidant enzyme activities. It thereby prevented lipid peroxidation and DNA damage, which was demonstrated by the inhibition of the formation of thiobarbituric acid reactive substance (TBARS), inhibition of the comet tail and decreased phospho-H2A.X expression. Annphenone protected Chinese hamster lung fibroblast (V79-4) cells from cell death via the inhibition of apoptosis induced by hydrogen peroxide (H(2)O(2)), as shown by decreased apoptotic nuclear fragmentation, decreased sub-G(1) cell population and inhibited mitochondrial membrane potential (Deltapsi) loss. Taken together, these findings suggest that annphenone exhibits antioxidant properties by inhibiting ROS generation and thus protecting cells from H(2)O(2)-induced cell damage.     

The mechanisms of oxidative DNA damage and apoptosis induced by norsalsolinol, an endogenous tetrahydroisoquinoline derivative associated with Parkinson's disease

Tetrahydroisoquinoline (TIQ) derivatives are putative neurotoxins that may contribute to the degeneration of dopaminergic neurons in Parkinson's disease. One TIQ, norsalsolinol (NorSAL), is present in dopamine-rich areas of human brain, including the substantia nigra. Here, we demonstrate that NorSAL reduces cell viability and induces apoptosis via cytochrome c release and caspase 3 activation in SH-SY5Y human neuroblastoma cells. Cytochrome c release, caspase 3 activation, and apoptosis induction were all inhibited by the antioxidant N -acetylcysteine. Thus, reactive oxygen species (ROS) contribute to apoptosis induced by NorSAL. Treatment with NorSAL also increased levels of oxidative damage to DNA, a stimulus for apoptosis, in SH-SY5Y. To clarify the mechanism of intracellular DNA damage, we examined the DNA damage caused by NorSAL using ³²P-5'-end-labeled isolated DNA fragments. NorSAL induced DNA damage in the presence of Cu(II). Catalase and bathocuproine, a Cu(I) chelator, inhibited this DNA damage, suggesting that ROS such as the Cu(I)-hydroperoxo complex derived from the reaction of H₂O₂ with Cu(I), promote DNA damage by NorSAL. In summary, NorSAL-generated ROS induced oxidative DNA damage, which led to caspase-dependent apoptosis in neuronal cells.     

镉、汞暴露在中华绒螯蟹卵巢中的富集及其氧化应激反应

研究以中华绒螯蟹(Eriocheir sinensis)为实验对象, 用不同浓度的Cd2+ 溶液(0、0.63、1.26、2.52、5.04和10.07 mg/L)或Hg2+ 溶液(0、0.06、0.11、0.23、0.46和0.91 mg/L)对其分别进行染毒, 6d后解剖取其卵巢, 检测卵巢中Cd和Hg的富集量以及氧化应激相关指标的变化情况。结果显示, 随着Cd2+或Hg2+染毒浓度的升高, 卵巢中Cd和Hg的富集量显著增加; H₂O₂、MDA和NO含量、MT mRNA表达量及NOS活性均随着Cd2+和Hg2+浓度的增加出现上升的趋势, 但SOD、CAT和GPx活性呈先升后降的趋势。这说明镉、汞均可引起卵巢组织产生过量的自由基和脂质过氧化, 并诱导氧化应激酶及MT mRNA的表达以抵抗不良的环境。总之, MT对中华绒螯蟹卵巢中重金属的解毒发挥着重要作用, 抗氧化酶能有效地消除活性氧自由基(ROS), 维持机体的氧化还原处于平衡状态, 共同保护膜脂免受氧化损伤。     

Influence of novel naphthalimide-based organoselenium on genotoxicity induced by an alkylating agent: the role of reactive oxygen species and selenoenzymes

Abstract

Objective

The protection conferred by a series of synthetic organoselenium compounds against genotoxicity and oxidative stress induced by a reference mutagen cyclophosphamide (CP) was assessed.

Method

Genotoxicity was induced in mice by CP treatment (25 mg/kg b.w.) for 10 consecutive days. Organoselenium compounds (3 mg/kg b.w.) were administered orally in a concomitant and pretreatment schedule. DNA damage in peripheral blood lymphocytes and frequency of chromosomal aberration in the bone marrow cells were measured. Liver tissues were collected for analysis of the activity of antioxidant and detoxifying enzymes, lipid peroxidation (LPO) level, glutathione content, and histopathology.

Results

Exposure to CP not only led to a significant increase in the percent of chromosomal aberration and DNA damage, but also enhanced generation of hepatic reactive oxygen species (ROS) and LPO level. The organoselenium compounds demonstrated marked functional protection against CP-induced genotoxicity. DNA damage and chromosomal aberration along with ROS generation were attenuated in the organoselenium-treated mice compared with the CP-treated control mice. CP caused marked depression in the activities of the selenoenzymes (glutathione peroxidase (GPx) and thioredoxin reductase (TRxR)) and other detoxifying and antioxidant enzymes, while treatment with organoselenium compounds restored all these activities towards normal.

Discussion

The protective effect of these compounds may be primarily associated with the improvement of the activity of antioxidant and detoxifying enzymes (including the selenoenzymes, GPx, and TRxR) that are known to protect the DNA and other cellular components from oxidative damage.     

Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems

Free radicals derived from oxygen, nitrogen and sulphur molecules in the biological system are highly active to react with other molecules due to their unpaired electrons. These radicals are important part of groups of molecules called reactive oxygen/nitrogen species (ROS/RNS), which are produced during cellular metabolism and functional activities and have important roles in cell signalling, apoptosis, gene expression and ion transportation. However, excessive ROS attack bases in nucleic acids, amino acid side chains in proteins and double bonds in unsaturated fatty acids, and cause oxidative stress, which can damage DNA, RNA, proteins and lipids resulting in an increased risk for cardiovascular disease, cancer, autism and other diseases. Intracellular antioxidant enzymes and intake of dietary antioxidants may help to maintain an adequate antioxidant status in the body. In the past decades, new molecular techniques, cell cultures and animal models have been established to study the effects and mechanisms of antioxidants on ROS. The chemical and molecular approaches have been used to study the mechanism and kinetics of antioxidants and to identify new potent antioxidants. Antioxidants can decrease the oxidative damage directly via reacting with free radicals or indirectly by inhibiting the activity or expression of free radical generating enzymes or enhancing the activity or expression of intracellular antioxidant enzymes. The new chemical and cell-free biological system has been applied in dissecting the molecular action of antioxidants. This review focuses on the research approaches that have been used to study oxidative stress and antioxidants in lipid peroxidation, DNA damage, protein modification as well as enzyme activity, with emphasis on the chemical and cell-free biological system.     

Zearalenone induces ROS‐mediated mitochondrial damage in porcine IPEC‐J2 cells

In this study, the mitochondrial damage effect and mechanism of zearalenone (ZEA) in swine small intestine IPEC‐J2 cells in vitro were comprehensively characterized. The analyses revealed that ZEA at high doses (8 and 7 μg/mL) can significantly increase P < 0.05 the malondialdehyde levels and decrease antioxidant enzymes activities after 48 h of exposure. Meanwhile, the reactive oxygen species (ROS) accumulation increased in high dose ZEA‐treated groups after 2 h treatment, but decreased due to the ROS‐induced mitochondrial damage and the caused cell apoptosis after 48 h of high does ZEA treatment. Moreover, the decreasing of mitochondrial membrane potential (MMP; ΔΨ) in high dose ZEA exposure was observed in line with the increasing ROS production in mitochondria. Results suggest that ZEA exposure can induce mitochondrial damage by reducing antioxidant enzyme activities, accumulation of ROS, and decreasing MMP. The mitochondrial damage had a dramatic concentration–effects relationship with ZEA.     

Effects of naringin on cytosine arabinoside (Ara-C)-induced cytotoxicity and apoptosis in P388 cells

Naringin (NG), a flavonoid in grapefruit and citrus, has been reported to exhibit antioxidant effects and pharmacological actions. Recently, we have reported that NG suppressed the cytotoxicity and apoptosis induced by H(2)O(2), a typical pro-oxidant, in mouse leukemia P388 cells. Cytosine arabinoside (1-beta-d-arabinofuranosylcytosine; Ara-C) is the most important antimetabolite chemotherapeutic drug used for acute leukemia. It has been suggested that Ara-C-induced cytotoxicity is caused by apoptosis, which is mediated by reactive oxygen species (ROS). In this study, we examined the effect of NG on the cytotoxicity and apoptosis in mouse leukemia P388 cells treated with Ara-C. Ara-C caused cytotoxicity in a concentration and time-dependent manner in the cells. N-Acetyl-L-cysteine (NAC), cystamine (CysA) or a reduced form of glutathione (GSH), typical antioxidants significantly blocked Ara-C-induced cytotoxicity. Similarly, Ara-C-induced cell death was completely prevented by NG. NG strongly reduced ROS production caused by Ara-C in the cells. NG slightly increased the activities of antioxidant enzymes, catalase and glutathione peroxidase. Ara-C caused apoptosis with nuclear morphological change and DNA fragmentation. NG remarkably attenuated the Ara-C-induced apoptosis. NG completely blocked the DNA damage caused by Ara-C treatment at 6 h using the Comet assay. Our data suggest that NG reduces Ara-C-induced oxidative stress through both an inhibition of the generation of ROS production and an increase in antioxidant enzyme activities. Consequently, NG blocked apoptosis caused by Ara-C-induced oxidative stress, resulting in the inhibition of the cytotoxicity of Ara-C.     

诺丽叶片的离体再生

以诺丽(Morinda citrifolia)叶片为外植体,在添加不同激素种类和浓度的MS培养基上进行离体培养,建立两种离体再生模式:模式Ⅰ为先脱分化愈伤组织,再分化不定根和不定芽;模式Ⅱ为直接培养生根后分化不定芽。结果表明:在模式Ⅰ中,诱导诺丽叶片产生愈伤组织的最优培养基为MS+0.1 mg·L~(-1)6-BA+2.0mg·L~(-1)2,4-D;诱导叶片愈伤组织再分化出不定根和不定芽的最优培养基为MS+1.0 mg·L~(-1)6-BA+0.4 mg·L~(-1) NAA或MS+2.0 mg·L~(-1)6-BA+0.4 mg·L~(-1) NAA,其中MS+1.0 mg·L~(-1)6-BA+0.4 mg·L~(-1) NAA生根时间最早为10 d左右,根系较发达,而MS+2.0 mg·L~(-1)6-BA+0.4 mg·L~(-1) NAA生根时间在15 d左右,根系发达。在模式Ⅱ中,诱导叶片直接生根长芽的培养基为MS+1.0 mg·L~(-1)6-BA+0.4 mg·L~(-1) NAA。将模式Ⅰ和模式Ⅱ中,完成诺丽叶片离体再生的苗切下后接种到MS+0.2 mg·L~(-1) NAA培养基中诱导生根,15 d左右分化出不定根,45 d获得完整植株。该研究结果为后续的遗传转化和基因改良研究奠定了基础。